Zhang S-X, Shan W-X, Yuan L-P, Liu Y-L, Sun L-Z
Department of Obstetrics and Gynecology, the First People's Hospital of Xuzhou, Xuzhou, Jiangsu, China.
Eur Rev Med Pharmacol Sci. 2016 Jul;20(13):2835-41.
We investigated the effects of small interference RNA (siRNA) on the cell proliferation inhibition, sensitivity to radiotherapy effects and cell apoptosis. The siRNA used here was specific to the pituitary tumor transforming gene (PTTG).
Vectors containing the specific functional siRNAs for PTTG were designed and constructed. Cells were divided into four groups: (I) blank control group; (II) radiotherapy group: cells were exposed to X-ray radiation; (III) Group PTTG siRNA: transfected with PTTG siRNA; (IV) PTTG siRNA+ radiotherapy group: transfected with PTTG siRNA and then were exposed to X-ray radiation. HEC-1A cells were transfected by the specific interfering plasmids using Lipofectamine 2000 transfection reagent. The PTTG protein expression levels were analyzed using Western blot Cell proliferation was examined by MTT assay and the HEC-1A cell line apoptosis was evaluated by flow cytometry.
Recombinant small interference RNA (siRNA) expression vectors targeting PTTG were successfully constructed. The results of MTT showed that the growth of the HEC-1A cell was negatively influenced after cells were transfected with PTTG siRNA. Furthermore, PTTG siRNA combined with radiotherapy demonstrated more powerful inhibitory effects. Cell apoptosis rates were significantly increased in the radiotherapy group and the PTTG siRNA transfection group when compared to the control group. A more pronounced cell apoptosis rate was observed in the group that was treated with PTTG siRNA combined with radiotherapy.
Recombinant small interference RNA (siRNA) expression vector targeting PTTG successfully inhibited the cell proliferation and induced apoptosis in endometrial carcinoma cells and increased the cancer cells vulnerability to the effects of radiation.
我们研究了小干扰RNA（siRNA）对细胞增殖抑制、放疗效果敏感性及细胞凋亡的影响。此处使用的siRNA对垂体肿瘤转化基因（PTTG）具有特异性。
设计并构建了含有针对PTTG的特定功能性siRNA的载体。细胞分为四组：（I）空白对照组；（II）放疗组：细胞接受X射线辐射；（III）PTTG siRNA组：转染PTTG siRNA；（IV）PTTG siRNA + 放疗组：转染PTTG siRNA后再接受X射线辐射。使用Lipofectamine 2000转染试剂通过特定干扰质粒转染HEC - 1A细胞。采用蛋白质免疫印迹法分析PTTG蛋白表达水平，通过MTT法检测细胞增殖，并通过流式细胞术评估HEC - 1A细胞系凋亡情况。
成功构建了靶向PTTG的重组小干扰RNA（siRNA）表达载体。MTT结果显示，转染PTTG siRNA后HEC - 1A细胞的生长受到负面影响。此外，PTTG siRNA联合放疗显示出更强的抑制作用。与对照组相比，放疗组和PTTG siRNA转染组的细胞凋亡率显著增加。PTTG siRNA联合放疗处理的组观察到更明显的细胞凋亡率。
靶向PTTG的重组小干扰RNA（siRNA）表达载体成功抑制了子宫内膜癌细胞的增殖并诱导凋亡，增加了癌细胞对辐射作用的敏感性。
